A below-ground herbivore shapes root defensive chemistry in natural plant populations

Plants display extensive intraspecific variation in secondary metabolites. However, the selective forces shaping this diversity remain often unknown, especially below ground. Using Taraxacum officinale and its major native insect root herbivore Melolontha melolontha, we tested whether below-ground herbivores drive intraspecific variation in root secondary metabolites. We found that high M. melolontha infestation levels over recent decades are associated with high concentrations of major root latex secondary metabolites across 21 central European T. officinale field populations. By cultivating offspring of these populations, we show that both heritable variation and phenotypic plasticity contribute to the observed differences. Furthermore, we demonstrate that the production of the sesquiterpene lactone taraxinic acid β-d-glucopyranosyl ester (TA-G) is costly in the absence, but beneficial in the presence of M. melolontha, resulting in divergent selection of TA-G. Our results highlight the role of soil-dwelling insects for the evolution of plant defences in nature.

Projected near-future CO2 levels increase activity and alter defensive behaviours in the tropical squid Idiosepius pygmaeus

Carbon dioxide (CO2) levels projected to occur in the oceans by the end of this century cause a range of behavioural effects in fish, but whether other highly active marine organisms, such as cephalopods, are similarly affected is unknown. We tested the effects of projected future CO2 levels (626 and 956 µatm) on the behaviour of male two-toned pygmy squid, Idiosepius pygmaeus. Exposure to elevated CO2 increased the number of active individuals by 19-25% and increased movement (number of line-crosses) by nearly 3 times compared to squid at present-day CO2. Squid vigilance and defensive behaviours were also altered by elevated CO2 with >80% of individuals choosing jet escape responses over defensive arm postures in response to a visual startle stimulus, compared with 50% choosing jet escape responses at control CO2. In addition, more escape responses were chosen over threat behaviours in body pattern displays at elevated CO2 and individuals were more than twice as likely to use ink as a defence strategy at 956 µatm CO2, compared with controls. Increased activity could lead to adverse effects on energy budgets as well as increasing visibility to predators. A tendency to respond to a stimulus with escape behaviours could increase survival, but may also be energetically costly and could potentially lead to more chases by predators compared with individuals that use defensive postures. These results demonstrate that projected future ocean acidification affects the behaviours of a tropical squid species.

A model of economic management of cultural heritage: the rehabilitation of fort christmas as an interpretation center of defensive architecture of the Mediterranean in Cartagena (Spain)

The Cultural Heritage constitutes a way to generate social identities and play an important role in the development of the Spanish Mediterranean cities that opt to sustainable quality tourism. The reflection on the necessity of intervention on this heritage, in addition to establishing what should be done, brings up the need to define the reasons for taking action, why and what-for. These decisions are essential to establish if its maintenance and recovery are economically sustainable. The Project "Cartagena Port of Cultures", with support from the European Union, is an example of effective instrument for ensuring the sustainability of our built heritage conservation. Its main objective was to enable sustainable development of tourism in Cartagena based on sustainability and seasonality. This was achieved through a process of recovery of heritage resources and their optimum promotion and marketing.

Effects of coaches' timeouts on basketball teams' offensive and defensive performances according to momentary differences in score and game period.

The aim of the present study was to assess the effects of game timeouts on basketball teams? offensive and defensive performances according to momentary differences in score and game period. The sample consisted of 144 timeouts registered during 18 basketball games randomly selected from the 2007 European Basketball Championship (Spain). For each timeout, five ball possessions were registered before (n?493) and after the timeout (n?475). The offensive and defensive efficiencies were registered across the first 35 min and last 5 min of games. A k-means cluster analysis classified the timeouts according to momentary score status as follows: losing ( ?10 to ?3 points), balanced ( ?2 to 3 points), and winning (4 to 10 points). Repeated-measures analysis of variance identified statistically significant main effects between pre and post timeout offensive and defensive values. Chi-square analysis of game period identified a higher percentage of timeouts called during the last 5 min of a game compared with the first 35 min (64.999.1% vs. 35.1910.3%; x ?5.4, PB0.05). Results showed higher post timeout offensive and defensive performances. No other effect or interaction was found for defensive performances. Offensive performances were better in the last 5 min of games, with the least differences when in balanced situations and greater differences when in winning situations. Results also showed one interaction between timeouts and momentary differences in score, with increased values when in losing and balanced situations but decreased values when in winning situations. Overall, the results suggest that coaches should examine offensive and defensive performances according to game period and differences in score when considering whether to call a timeout.

Routines in precision agriculture technology: an action research study

A crop management and precision agriculture software application facilitates the flow of information between disparate software/equipment and the network of individuals that work with them. A new generation of farmers are using precision technology to help them more efficiently manage their roplands. By measuring precisely the way their fields reflect and emit energy at visible and infrared wavelengths, precision farmers can monitor a wide range of variables that affect their crops,such as soil moisture, surface temperature, photosynthetic activity, and weed or pest infestations. Over thirty years have passed since Nelson and Winter put the concept of routines firmly at the center of the analysis of organizational and economic change. Taken as the central unit of analysis, routines would help understand energy and agriculture economy evolution.

Self-organized defensive behavior in honeybees

We investigated the defensive behavior of honeybees under controlled experimental conditions. During an attack on two identical targets, the spatial distribution of stings varied as a function of the total number of stings, evincing the classic “pitchfork bifurcation” phenomenon of nonlinear dynamics. The experimental results support a model of defensive behavior based on a self-organizing mechanism. The model helps to explain several of the characteristic features of the honeybee defensive response: (i) the ability of the colony to localize and focus its attack, (ii) the strong variability between different hives in the intensity of attack, as well as (iii) the variability observed within the same hive, and (iv) the ability of the colony to amplify small differences between the targets.

Firefly “femmes fatales” acquire defensive steroids (lucibufagins) from their firefly prey

Female fireflies of the genus Photuris, the so-called firefly “femmes fatales,” prey on male fireflies of the genus Photinus. The females are able to entrap the males by faking the flash signal characteristics of the Photinus female. We found that by feeding on Photinus males, Photuris females gain more than nutrients. They also acquire defensive steroidal pyrones called lucibufagins, which are contained in Photinus but which Photuris fireflies are unable to produce on their own. Photuris females that eat Photinus males or lucibufagin are rejected by Phidippus jumping spiders. Lucibufagin itself proved to be a deterrent to such spiders. Field-collected Photuris females contain lucibufagin in varying amounts. The more lucibufagin they contain the more unacceptable they are to Phidippus.

Miriamin, a defensive diterpene from the eggs of a land slug (Arion sp.)

The eggs of the land slug Arion sp. contain a diterpene, miriamin, characterized as a polyoxygenated geranylgeraniol derivative. In bioassays with a coccinellid beetle, Harmonia axyridis, miriamin was shown to be potently antifeedant, indicating that the compound plays a protective role in nature. It is suggested that mucilaginous soil-inhabiting organisms, given their intense exposure to pathogens and predators, may be a rich source of chemical defensive agents.

Defensive extrusive ectosymbionts of Euplotidium (Ciliophora) that contain microtubule-like structures are bacteria related to Verrucomicrobia

Epixenosomes, ectosymbionts on hypotrich ciliates (genus Euplotidium) defend their host against the ciliate predator Litonotus lamella. Although here only Euplotidium itoi and Euplotidium arenarium from tide pools along a rocky shore near Leghorn (Ligurian sea) were studied in detail, these epibionts are certainly present on specimens of E. itoi and on other Euplotidium species in similar north coastal habitats. The complex life history of epixenosomes has two main stages. In stage I, cells with typical prokaryotic structure divide by binary fission. Stage II cells show complex organization with different cytoplasmic compartments where an extrusive apparatus within a proteinaceous matrix, although not membrane-bounded, differs from the remaining cytoplasm. The ejection process is involved in defense; extrusive apparatus is surrounded by a basket consisting of bundles of tubules. These tubules, 22 ± 3 nm in diameter, delimited by a wall made up of globular structures, are sensitive to inhibitor of tubulin polymerization (nocodazole/4°C temperature) and react positively with different antitubulin antibodies, two of which are monoclonal. The prokaryotic vs. eukaryotic nature of epixenosomes was resolved by comparative sequence analysis of amplified small subunit rRNA genes and in situ hybridization with fluorescently labeled rRNA-targeted polynucleotide probes. These unique ectosymbionts are phylogenetically related to Verrucomicrobia. Epixenosomes represent marine symbionts in this recently discovered division of the Bacteria.

Chickpea Defensive Proteinase Inhibitors Can Be Inactivated by Podborer Gut Proteinases1

Developing chickpea (Cicer arietinum L.) seeds 12 to 60 d after flowering (DAF) were analyzed for proteinase inhibitor (Pi) activity. In addition, the electrophoretic profiles of trypsin inhibitor (Ti) accumulation were determined using a gel-radiographic film-contact print method. There was a progressive increase in Pi activity throughout seed development, whereas the synthesis of other proteins was low from 12 to 36 DAF and increased from 36 to 60 DAF. Seven different Ti bands were present in seeds at 36 DAF, the time of maximum podborer (Helicoverpa armigera) attack. Chickpea Pis showed differential inhibitory activity against trypsin, chymotrypsin, H. armigera gut proteinases, and bacterial proteinase(s). In vitro proteolysis of chickpea Ti-1 with various proteinases generated Ti-5 as the major fragment, whereas Ti-6 and -7 were not produced. The amount of Pi activity increased severalfold when seeds were injured by H. armigera feeding. In vitro and in vivo proteolysis of the early- and late-stage-specific Tis indicated that the chickpea Pis were prone to proteolytic digestion by H. armigera gut proteinases. These data suggest that survival of H. armigera on chickpea may result from the production of inhibitor-insensitive proteinases and by secretion of proteinases that digest chickpea Pis.

Polypeptide signaling for plant defensive genes exhibits analogies to defense signaling in animals.

The activation of plant defensive genes in leaves of tomato plants in response to herbivore damage or mechanical wounding is mediated by a mobile 18-amino acid polypeptide signal called systemin. Systemin is derived from a larger, 200-amino acid precursor called prosystemin, similar to polypeptide hormones and soluble growth factors in animals. Systemin activates a lipid-based signaling cascade, also analogous to signaling systems found in animals. In plants, linolenic acid is released from membranes and is converted to the oxylipins phytodienoic acid and jasmonic acid through the octadecanoid pathway. Plant oxylipins are structural analogs of animal prostaglandins which are derived from arachidonic acid in response to various signals, including polypeptide factors. Constitutive overexpression of the prosystemin gene in transgenic tomato plants resulted in the overproduction of prosystemin and the abnormal release of systemin, conferring a constitutive overproduction of several systemic wound-response proteins (SWRPs). The data indicate that systemin is a master signal for defense against attacking herbivores. The same defensive proteins induced by wounding are synthesized in response to oligosaccharide elicitors that are generated in leaf cells in response to pathogen attacks. Inhibitors of the octadecanoid pathway, and a mutation that interrupts this pathway, block the induction of SWRPs by wounding, systemin, and oligosaccharide elicitors, indicating that the octadecanoid pathway is essential for the activation of defense genes by all of these signals. The tomato mutant line that is functionally deficient in the octadecanoid pathway is highly susceptible to attacks by Manduca sexta larvae. The similarities between the defense signaling pathway in tomato leaves and those of the defense signaling pathways of macrophages and mast cells of animals suggests that both the plant and animal pathways may have evolved from a common ancestral origin.

Oligogalacturonides and chitosan activate plant defensive genes through the octadecanoid pathway.

Jasmonic acid, synthesized from linolenic acid (the octadecanoid pathway), has been proposed to be part of a signal transduction pathway that mediates the induction of defensive genes in plants in response to oligouronide and polypeptide signals generated by insect and pathogen attacks. We report here that the induction of proteinase inhibitor accumulation in tomato leaves by plant-derived oligogalacturonides and fungal-derived chitosan oligosaccharides is severely reduced by two inhibitors (salicylic acid and diethyldi-thiocarbamic acid) of the octadecanoid pathway, supporting a role for the pathway in signaling by oligosaccharides. Jasmonic acid levels in leaves of tomato plants increased several fold within 2 hr after supplying the polypeptide systemin, oligogalacturonides, or chitosan to the plants through their cut stems, as expected if they utilize the octadecanoid pathway. The time course of jasmonic acid accumulation in tomato leaves in response to wounding was consistent with its proposed role in signaling proteinase inhibitor mRNA and protein synthesis. The cumulative evidence supports a model for the activation of defensive genes in plants in response to insect and pathogen attacks in which various elicitors generated at the attack sites activate the octadecanoid pathway via different recognition events to induce the expression of defensive genes in local and distal tissues of the plants.

Comparison of the defensive elicitation activity of cerato-populin and cerato-platanin: a structural model

Plants can defend themselves from potential pathogenic microorganisms relying on a complex interplay of signaling pathways: activation of the MAPK cascade, transcription of defense related genes, production of reactive oxygen species, nitric oxide and synthesis of other defensive compounds such as phytoalexins. These events are triggered by the recognition of pathogen’s effectors (effector-triggered immunity) or PAMPs (PAMP-triggered immunity). The Cerato Platanin Family (CPF) members are Cys-rich proteins secreted and localized on fungal cell walls, involved in several aspects of fungal development and pathogen-host interactions. Although more than hundred genes of the CPF have been identified and analyzed, the structural and functional characterization of the expressed proteins has been restricted only to few members of the family. Interestingly, those proteins have been shown to bind chitin with diverse affinity and after foliar treatment they elicit defensive mechanisms in host and non-host plants. This property turns cerato platanins into interesting candidates, worth to be studied to develop new fungal elicitors with applications in sustainable agriculture. This study focus on cerato-platanin (CP), core member of the family and on the orthologous cerato-populin (Pop1). The latter shows an identity of 62% and an overall homology of 73% with respect to CP. Both proteins are able to induce MAPKs phosphorylation, production of reactive oxygen species and nitric oxide, overexpression of defense’s related genes, programmed cell death and synthesis of phytoalexins. CP, however, when compared to Pop1, induces a faster response and, in some cases, a stronger activity on plane leaves. Aim of the present research is to verify if the dissimilarities observed in the defense elicitation activity of these proteins can be associated to their structural and dynamic features. Taking advantage of the available CP NMR structure, Pop1’s 3D one was obtained by homology modeling. Experimental residual dipolar couplings and 1H, 15N, 13C resonance assignments were used to validate the model. Previous works on CPF members, addressed the highly conserved random coil regions (loops b1-b2 and b2-b3) as sufficient and necessary to induce necrosis in plants’ leaves: that region was investigated in both Pop1 and CP. In the two proteins the loops differ, in their primary sequence, for few mutations and an insertion with a consequent diversification of the proteins’ electrostatic surface. A set of 2D and 3D NMR experiments was performed to characterize both the spatial arrangement and the dynamic features of the loops. NOE data revealed a more extended network of interactions between the loops in Pop1 than in CP. In addition, in Pop1 we identified a salt bridge Lys25/Asp52 and a strong hydrophobic interaction between Phe26/Trp53. These structural features were expected not only to affect the loops’ spatial arrangement, but also to reduce the degree of their conformational freedom. Relaxation data and the order parameter S2 indeed highlighted reduced flexibility, in particular for loop b1-b2 of Pop1. In vitro NMR experiments, where Pop1 and CP were titrated with oligosaccharides, supported the hypothesis that the loops structural and dynamic differences may be responsible for the different chitin-binding properties of the two proteins: CP selectively binds tetramers of chitin in a shallow groove on one side of the barrel defined by loops b1-b2, b2-b3 and b4-b5, Pop1, instead, interacts in a non-specific fashion with oligosaccharides. Because the region involved in chitin-binding is also responsible for the defense elicitation activity, possibly being recognized by plant's receptors, it is reasonable to expect that those structural and dynamic modifications may also justify the different extent of defense elicitation. To test that hypothesis, the initial steps of a protocol aimed to the identify a receptor for CP, in silico, are presented.

Phraseological features in conversational routines: An integrative model of autonomy

This paper deals with the analysis of routine formulas according to their degree of independence in discourse, with the aim of establishing a gradual line which restructures Sphere III (Corpas 1996, Alvarado 2015) — where phraseological utterances belong. The system developed by Briz and the Val.Es.Co. Group (2003, 2014) will help us segment conversation and check that phraseological utterances show various degrees of independence, both in Peninsular Spanish and in Latin American Spanish, which will make it possible to restructure Sphere III. The methodology utilized corresponds to the phraseological and pragmatic approach, and the examples were extracted from the Corpus de Conversaciones Coloquiales of Briz and the Val.Es.Co. Group (2002), and from the Corpus Preseea (2014).

A Defensive "stand-by reaction" at Critical Moments of the Reproductive Life Cycle

Investigation about the psychological experiences of the reproductive life cycle showed that in critical moments special reactions may happen. These reactions seem to be defensive in nature, are set in motion in order to promote some kind of emotional protection and are performed in two opposite directions: a) a decreasing of the contact with aggressive impulses and b) an increasing of the use of rationalization and denial of frustrating situations. Examples of those rearrangements were observed at samples of: 1) pregnant women in obstetric high-risk consultation, 2) infertile couples waiting for infertility consultations and 3) pregnant women waiting for amniocentesis results. These data seem to be in accordance with the classical psychological points of view: a) gestation should be considered as a period of protection, b) during pregnancy a “primary maternal preoccupation” (Winnicot, 1958) emerges leading to the mobilization of all resources available for pregnant women and c) along gestational development psychological changes show how flexible maternal functioning may become. What was not expected is that in the absence of pregnancy, infertile couples should behave very similarly to what it is observed when pregnancy is in danger or when medical problems about the mother’s or the baby’s health arise in the horizon. Due to its “freezing” consequences upon emotional development we propose that this kind of reaction will be designated as “stand-by reaction”.